I am curious if anyone has used Pharmacia's pCANTAB5 for phagemid
expression of scFv. I am thinking of ordering the kit and want to
know how it works and if there are a lot of problems. It appears from
the way the vector is designed that you incorporate 4 aa in the
5' end of your scFv. It looks like, Ala, Ala, Gln, and either
arg, Pro, Leu, Gln,or His. Is this really how it works and does this
present a problem when studying the protein? Also, can you recover
soluble scFv protein to study from this kit or put the vector in
a non-suppressor strain and insert an amber codon and produce soluble
protein that way? Any feedback or experience with this system would
be appreciated.
Thanks
Cathy Rumbley
crumbley at uxa.cso.uiuc.edu